PT  - JOURNAL ARTICLE
AU  - Heidi Kristine Grønlien
AU  - Romain Fontaine
AU  - Kjetil Hodne
AU  - Isabelle Tysseng
AU  - Eirill Ager-Wick
AU  - Finn-Arne Weltzien
AU  - Trude Marie Haug
TI  - Gonadotropes in medaka grow long extensions with varicosity-like swellings, projecting towards each other and blood vessels
AID  - 10.1101/777151
DP  - 2019 Jan 01
TA  - bioRxiv
PG  - 777151
4099  - http://biorxiv.org/content/early/2019/09/20/777151.short
4100  - http://biorxiv.org/content/early/2019/09/20/777151.full
AB  - Accumulating evidence in the scientific literature indicates that some pituitary cell types are organized in complex networks. Previous observations have indicated that this may also be the case in medaka (Oryzias latipes), where long cellular extensions with varicosity-like swellings are formed by luteinizing hormone (Lh)-producing gonadotropes expressing green fluorescent protein. In this study, immunofluorescence of intact pituitaries reveal that Lh beta polypeptides are mainly located in the varicosity-like swellings and at the extremity of the extensions. Some extensions approach nearby Lh-producing cells, and other extensions are in close contact with blood vessels. To investigate whether these extensions may contribute to network formation, we followed their development using confocal and fluorescent microscopy on primary cultures. During the first two days in culture, the extensions initiated the formation of homotypic cellular networks and clustering. The extensions were classified as either major or minor. Major extensions were several cell diameters long, dependent on microtubules, and displaying varicosity-like swellings at regular intervals. Minor extensions typically protruded from the major, were significantly shorter and thinner, and dependent on actin. The swellings were dependent on both microtubules and actin. Flash photolysis of caged Ca2+ showed that the signal was propagated along the major extensions, intensifying in each swelling, indicating a continuous structure. However, the Ca2+ signal did not transfer to the next cell in the network, but was transferred between cells merged at their somas. In summary, Lh-producing gonadotropes in medaka display a complex cellular structure of extensions, possibly linked to communication with blood vessels and/or other gonadotrope cells.